Attachment of an NMR-invisible solubility enhancement tag using a sortase-mediated protein ligation method
Attachment of an NMR-invisible solubility enhancement tag using a sortase-mediated protein ligation method
复制标题
DOI：
10.1007/s10858-008-9296-5
复制
发表时间：
2009-03-01
影响因子：
2.7
通讯作者：
Inagaki, Fuyuhiko
中科院分区：
文献类型：
--
作者：
Kobashigawa, Yoshihiro;Kumeta, Hiroyuki;Inagaki, Fuyuhiko
样品的溶解度是必不可少的结构研究蛋白质的溶液核磁共振。将溶解度增强标签（如GB1、MBP和硫氧还蛋白）附着到靶蛋白上已被用于此目的。然而，标签与目标蛋白的信号重叠往往使光谱分析变得困难。在这里，我们报道了一种排序酶介导的蛋白质连接方法，通过在c端制备同位素标记的靶蛋白与未标记的GB1标签连接来消除标签产生的NMR信号。
Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Here we report a sortase-mediated protein ligation method to eliminate NMR signals arising from the tag by preparing the isotopically labeled target protein attached with the non-labeled GB1 tag at the C-terminus.